Knowing the host range of a pathogen is critical to developing and implementing effective disease management programs. Calonectria pseudonaviculata (Cps) is known to attack a number of species, varieties, and cultivars in the genus Buxus as well as three Pachysandra species (Pachysandra terminalis, Pachysandra procumbens, and Pachysandra axillaris) and several Sarcococca species, all in the Buxaceae family. The objective of this study was to evaluate non-Buxaceae groundcovers and companion plants commonly associated with boxwood plantings for their susceptibility to Cps. Twenty-seven plant species belonging to 21 families were exposed to different levels of inoculum: 50 to 300 conidia per drop for detached leaf assays and 30,000 to 120,000 conidia per 1 ml for whole-plant assays. Inoculated plants were incubated in humid environments for at least 48 h to facilitate infection. Cps infection and sporulation were observed on 12 plant species: Alchemilla mollis, Arctostaphylos uva-ursi, Brunnera macrophylla, Epimedium × youngianum, Galium odoratum, Geranium sanguineum, Phlox subulata, Tiarella cordifolia, Callirhoe involucrata, Iberis sempervirens, Mazus reptans, and Vinca minor. These results suggest that there may be more hosts of Cps commonly grown in nurseries and landscapes. If corroborated by observations of natural infection, these findings have implications for the Boxwood Blight Cleanliness Program instituted by the National Plant Board and for planning disease mitigation at production and in the landscape.

SSG, a Burkholderia isolate recovered from boxwood leaves, has many genes and gene clusters coding plant growth promoting traits and antagonistic activity against plant pathogens. It has been demonstrated that this bacterial isolate is a potent biofertilizer for boxwood and an effective biocontrol agent against boxwood blight. In this study we evaluated SSG against 21 additional pathogens including six bacteria, nine fungi and six Phytophthora species using dual culture assays, and against twelve diseases in planta under controlled environment. SSG suppressed all fungal and Phytophthora species, and five of the six Gram negative bacteria in dual culture. SSG provided the most consistent control to bacterial blight of geranium caused by Xanthomonas campestriscampestriscampestriscampestris at 41% to 58%, and to pansy leaf spot by Colletotrichum fnicticola at 30% to 32% across all three treatment timings from 1 day before inoculation (dbi) to 3 days after inoculation (dai). Among the six other diseases evaluated for multiple treatment timings, SSG provided the greatest control at 1 dbi to two diseases with hydrangea anthracnose by C. gloeosporioides at 67% and rhododendron Ramorum blight by Phytophhtora ramorum at 54%. The best SSG control of pansy gray mold by Botrytis cinerea was 67% at 1 wbi while that of tomato spotted wilt virus of impatiens was 53% at 2 wbi. When plants were treated with SSG only at one timing of 1 dbi, the best disease control was observed on Phytophthora blight of annual vinca by P. nicotianae at 95%, followed by pansy leaf spot by Alternaria tenuissima at 60%, Phytophthora blight of bell pepper by P. capsici at 56%, and Volutella blight of boxwood by Pseudonectria rouselliana at 44%. Further investigations into its efficacy for control of two root/crown rot diseases are warranted. Nevertheless, this study demonstrates that SSG is a promising broad-spectrum biocontrol agent for a variety of plant diseases caused by diverse pathogens.

Background Agricultural runoff recycling systems are manmade aquatic ecosystems of growing significance to global water sustainability, crop health, and production. This study investigated the seasonal dynamics of microbial community in a three-reservoir recycling irrigation system with a stepwise water flow and compared with that of an adjacent runoff-free stream. Runoff water from all production areas was captured in a sedimentation reservoir which overflowed to a transition reservoir then retention reservoir through a culvert. Stream water was pumped to replenish the reservoirs as needed during growing seasons. Results 16S rDNA PCR clone libraries of quarterly water samples from three reservoirs and one stream were sequenced, and 575 operational taxonomic units (OTUs) were identified and assigned to cyanobacteria, eukaryotic phytoplankton, and other bacteria. When compared to the stream, three reservoirs consistently had low microbial diversity. A distinct seasonal pattern of microbial community structure was observed for each reservoir and the stream. Stream was consistently dominated by other bacteria. Retention reservoir was dominated by cyanobacteria during the summer and fall and eukaryotic phytoplankton during the winter and spring. Sedimentation reservoir was dominated by cyanobacteria beginning in the spring but that dominance was altered when water was pumped from the stream from early spring to fall seasons. Transition reservoir had the greatest shift of microbial community structure, being dominated by other bacteria in the summer, cyanobacteria in the fall, and eukaryotic phytoplankton in the winter and spring. Water temperature and ammonium level were the two most important contributing factors to the seasonality of microbial community in these reservoirs. Conclusions The three recycling irrigation reservoirs consistently had lower microbial diversity and distinct community structure when compared to the stream. These reservoirs were typically dominated by cyanobacteria during warm seasons and eukaryotic phytoplankton during cool seasons. This seasonal pattern was altered when water was pumped from the stream. The cyanobacteria dominance was associated with rising water temperature and ammonium level. These results highlight the importance of preventing agricultural runoff from entering natural waterways and water resources and provide a useful framework for further investigations into the ecological processes of this emerging ecosystem.

Triplicate water samples were collected monthly from nine waterways [eight runoff containment basins (RCBs) and one stream] on four commercial ornamental plant nurseries from February to July, and from one RCB and nursery from April to October. Four RCBs, one per nursery, were actively used as an irrigation water source. Analysis was done for 18 water quality variables, including ammonium–nitrogen (NH 4 + –N), nitrate–nitrogen (NO 3 − –N), ortho phosphate–phosphorus (PO 4 –P), total-phosphorus (T-P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), aluminum (Al), boron (B), copper (Cu), iron (Fe), manganese (Mn), zinc (Zn), pH, total alkalinity (T-Alk), electrical conductivity (EC), and sodium (Na). The degree and rate of monthly change varied considerably between RCBs. Macronutrients generally increased at most nurseries in 1–2 months after fertilizer application particularly in three RCBs (MD21, VA11, and VA12), with levels of N- and P forms exceeding preferred criteria for irrigation water by June and July in VA11 and VA12. Micronutrients fluctuated less but did vary per RCB with the most monthly change occurring in MD21. Even though pH fluctuated, pH tended to remain alkaline or neutral to acidic respective of the RCB during the entire sample period. T-Alk tended to increase over the summer. EC primarily fluctuated in RCBs with high macronutrient levels. Although levels of N- and P forms were mostly suitable by irrigation water criteria, they were frequently above U.S. Environmental Protection Agency (USEPA) nutrient criteria for promoting healthy water environments of lakes and reservoirs, and are discussed.

Nine runoff containment basins (RCBs), used directly or indirectly for irrigating plants in ornamental plant nurseries, and one adjacent stream were sampled for water quality between Feb. and July 2013 in Maryland (MD), Mississippi (MS), and Virginia (VA). Triplicate water samples were taken monthly. Analysis was done for 18 water quality variables including nitrate-nitrogen (NO3−-N) and ammonium-nitrogen (NH4+-N), orthophosphate-phosphorus (PO4-P) and total-phosphorus (T-P), potassium, calcium, magnesium, sulfur, aluminum, boron (B), copper (Cu), iron (Fe), manganese, zinc (Zn), pH, total alkalinity (T-Alk), electrical conductivity (EC), and sodium. Additionally, 15 RCBs from 10 nurseries in Alabama (AL), Louisiana (LA), and MS were sampled in 2014 and 2016. Most prevalent correlations (P = 0.01) were between macronutrients, EC, B, Fe, and Zn, but none were prevalent across a majority of RCBs. Water quality parameter values were mostly present at low to preferred levels in all 25 waterways. Macronutrient levels were highest for a RCB that receives fertility from fertigation derived runoff. Water pH ranged from acidic to alkaline (>8). Results of this study show water quality in RCBs can be suitable for promoting plant health in ornamental plant nurseries, but also shows levels will vary between individual RCBs, therefore demonstrates need to verify water quality from individual water sources.

Recycling irrigation reservoirs (RIRs) are emerging aquatic environments of global significance to crop production, water conservation and environmental sustainability. This study characterized the diversity and population structure of cyanobacteria and other detected microbes in water samples from eight RIRs and one adjacent runoff-free stream at three ornamental crop nurseries in eastern (VA1 and VA3) and central (VA2) Virginia after cloning and sequencing the 16S rRNA gene targeting cyanobacteria and chloroplast of eukaryotic phytoplankton. VA1 and VA2 utilize a multi-reservoir recycling irrigation system with runoff channeled to a sedimentation reservoir which then overflows into transition and retention reservoirs where water was pumped for irrigation. VA3 has a single sedimentation reservoir which was also used for irrigation. A total of 208 operational taxonomic units (OTU) were identified from clone libraries of the water samples. Among them, 53 OTUs (358 clones) were cyanobacteria comprising at least 12 genera dominated by Synechococcus species; 59 OTUs (387 clones) were eukaryotic phytoplankton including green algae and diatoms; and 96 were other bacteria (111 clones). Overall, cyanobacteria were dominant in sedimentation reservoirs, while eukaryotic phytoplankton and other bacteria were dominant in transition/retention reservoirs and the stream, respectively. These results are direct evidence demonstrating the negative impact of nutrient-rich horticultural runoff, if not contained, on natural water resources. They also help in understanding the dynamics of water quality in RIRs and have practical implications. Although both single- and multi-reservoir recycling irrigation systems reduce the environmental footprint of horticultural production, the former is expected to have more cyanobacterial blooming, and consequently water quality issues, than the latter. Thus, a multi-reservoir recycling irrigation system should be preferred where feasible.

Recycling irrigation reservoirs (RIRs) are an emerging aquatic ecosystem and water resource of global significance. This study investigated the vertical distribution of water temperature, dissolved oxygen (DO), and pH in eight RIRs at two nurseries each in Virginia and Maryland from 2011 to 2014. Monomictic thermal stratification was observed from April to October in all RIRs, despite their shallow depths (0.75-3.89 m). The strongest stratification had a top-bottom temperature difference of 21.53 degrees C. The top-bottom temperature difference was positively correlated with water column depth, air temperature, and daily light integral (p < 0.05). Wind speed did not impact the thermal stratification, likely due to their relatively small surface areas. Thermal stratification affected the vertical distribution of DO and pH. The top-bottom differences in DO and pH were greater during stratification periods than nonstratification periods. Water pH in all RIRs was higher at the top than at the bottom with the greatest difference of 4.16 units. Discovery and characterization of thermal stratification in RIRs helps understand water quality dynamics in this novel ecosystem and promote safe and productive water reuse for irrigation. Specifically, water withdrawal depths should be adjusted according to variations in temperature, DO, and pH during the stratification and nonstratification periods to mitigate pathogen risk and improve water treatment efficacy and crop production.

Recycling irrigation reservoirs (RIRs) are an emerging aquatic ecosystem of critical importance, for conserving and protecting increasingly scarce water resources. Here, we compare water quality between runoff entrance and middle of four RIRs in nurseries in Virginia (VA) and Maryland (MD). Surface water temperature (T) and oxidation-reduction potential (ORP) were lower in the middle than at the entrance, while the trend was opposite for dissolved oxygen (DO), pH and chlorophyll a (Chla). The magnitude of these differences between the entrance and middle decreased with increasing depth. These differences were magnified by water stratification from April to October. Minimum differences were observed for electrical conductivity (EC), total dissolved solids (TDS) and turbidity (TUR). Cluster analyses were performed on water quality difference data to evaluate whether the differences vary with respect to reservoirs. Two clusters were formed with one consisting primarily of VA reservoirs, and the other consisting mostly of MD reservoirs in both years. Water quality in the middle and at the entrance of RIRs was expected to vary greatly because of runoff inflow. The two-point water quality differences observed here, although statistically significant, are not large enough to cause significant impact on crop health and productivity for most water quality parameters except pH. Additional analysis of outlet data shows that the range and magnitude of water quality difference between the middle and the outlet are comparable to those between the middle and entrance of RIRs. These results indicate that monitoring at a single point is sufficient to obtain reliable water quality estimates for most water quality parameters in RIRs except pH. This is important when considering the cost of labor and equipment necessary for documenting water quality in agricultural production systems. However, additional pH measurements are still necessary to make practical water quality management decisions.

Recycling irrigation reservoirs (RIRs) are an emerging aquatic ecosystem of global significance. Building upon the recent discovery of thermal stratification in these novel systems and its impacts on the vertical distribution of dissolved oxygen and pH, this study investigated the dynamics of chlorophyll a (CHLA), oxidation reduction potential (ORP), electrical conductivity (EC), and turbidity in the water columns of eight RIRs in the Mid-Atlantic region in USA over a 3-year period from 2011 to 2014. The vertical distribution of CHLA was associated with thermal stratification while ORP and EC were directly affected by thermal stratification. Specifically, CHLA concentration was greatest in the middle thermocline, followed by lower hypolimnion during the Stratification period. ORP level was higher at the surface than bottom with differences up to 505.67 my in Reservoir VA23. EC level was higher in the bottom than at the surface with the maximum difference of 0.90 dS m(-1) in Reservoir VA22. The top-bottom ORP and EC differences increased when water was stratified. Turbidity was indirectly affected by stratification. The correlation of turbidity with total precipitation was stronger during the non-stratification period from November to March than during the stratification period from April to October, mainly due to restriction on water mixing. The characterization of these additional water quality parameters in the water columns of RIRs provides fundamental knowledge about this novel water system and will help in developing sound water quality management practices for improved crop health and production while reducing agriculture's environmental footprint. This study also provides additional evidence supporting pH as a significant indicator for monitoring water quality in RIRs. (C) 2015 Elsevier B.V. All rights reserved.

A novel Phytophthora species was frequently recovered from irrigation reservoirs at several ornamental plant production facilities in eastern Virginia. Initial sequencing of the internal transcribed spacer (ITS) region of this species generated unreadable sequences due to continual polymorphic positions. Cloning and sequencing the ITS region as well as sequencing the mitochondrially encoded cytochrome c oxidase 1 and beta-tubulin genes revealed that it is a hybrid between P. taxon PgChlamydo as its paternal parent and an unknown species genetically close to P. mississippiae as its maternal parent. This hybrid has some diagnostic morphological features of P. taxon PgChlamydo and P. mississippiae. It produces catenulate hyphal swellings, characteristic of P. mississippiae, and chlamydospores, typical of P. taxon PgChlamydo. It also produces both ornamented and relatively smooth-walled oogonia. Ornamented oogonia are another important diagnostic character of P. mississippiae. The relatively smooth-walled oogonia may be indicative of oogonial character of P. taxon PgChlamydo. The new hybrid is described here as Phytophthora ×stagnum.

In the summer of 2011, severe root and stem rot of begonia (Begonia × semperflorens-cultorum cv. Vodka Dark Red) was observed during a field trial. Seventy-eight percent of the plants had symptoms included foliar blight, blackened and rotting roots, rotting stems, and collapsing crown, often leading to plant death. Isolation from the diseased plant roots consistently recovered a Pythium-like species and 41 isolates were subcultured for identification. These isolates produced very similar single-strand conformation polymorphism (SSCP) fingerprints (2), which were distinct from those of other oomycete pathogens known to attack begonia (1). These isolates produced proliferous, ovoid to globose, terminal, and papillate sporangia which were 30.6 to 45.4 μm (av. 38.7 μm) in length and 20.5 to 35.4 μm (av. 28.2 μm) in width. Oogonia were produced in single culture grown in clarified V8 juice agar. These smooth-walled oogonia were mostly aplerotic and 28.9 to 36.8 μm (av. 33.1 μm) in diameter. Each contained a single oospore with a diameter of 23.7 to 34.4 μm (av. 26.9 μm). Single to multiple antheridia were attached lengthwise to each oogonium. These morphological characteristics match the description of Phytopythium helicoides (= Pythium helicoides) (3). The identity of these isolates was confirmed by sequencing the rDNA internal transcribed spacer (ITS) 1 and 2 regions. ITS sequence of the representative isolate 55C7 (GenBank Accession No. KC907734) had 97 to 99% homology with P. helicoides sequences in GenBank. Two isolates, 55C7 and 56A7, were tested for pathogenicity to begonia in the summer of 2012. Twelve plants per isolate were inoculated by injecting ground P. helicoides-colonized rice grains into the root soil using a long-neck funnel. Sterile rice grains were used on control plants. Aboveground symptoms including foliar blight, stem rot, and collapsing crown were observed 7 days after inoculation and the disease progressed for additional 6 weeks. At 7 weeks, all inoculated plants showed different symptom levels. Four and 10 plants inoculated with 55C7 and 56A7, respectively, were already dead. Begonia roots showed severe symptoms including blackening, stunted growth, and rotting. Seven of 12 control plants also had notable symptoms due to cross contamination. Isolates recovered from all symptomatic plants had identical SSCP fingerprints to those of isolates 55C7 and 56A7. To our knowledge, this is the first report of P. helicoides attacking begonia plants. The avenue of this pathogen entering the 2011 field trial remains unknown. The field trial in 2011 and pathogenicity test in 2012 indicate that this pathogen is potentially destructive to begonia. Additional research is warranted to identify the origin and dissemination of this pathogen to mitigate the risk to begonia production. References: (1) C. X. Hong et al. Plant Dis. 92: 1201, 2008. (2) P. Kong et al. FEMS Microbiol. Lett. 240:229, 2003. (3) A. J. van der Plaats-Niterink. Monograph of the Genus Pythium. Centraalbureau voor Schimmelcultures, Baarn, the Netherlands, 1981.

Two distinct subgroups (L2 and A(-2)) were recovered from irrigation reservoirs and a stream in Virginia, USA. After molecular, morphological and physiological examinations, the L2 subgroup was named Phytophthora aquimorbida and the A(-2) designated as Phytophthora taxon 'aquatilis'. Both taxa are homothallic. P. aquimorbida is characterized by its noncaducous and nonpapillate sporangia, catenulate and radiating hyphal swellings and thick-walled plerotic oospores formed in globose oogonia mostly in the absence of an antheridium. P. taxon 'aquatilis' produces plerotic oospores in globose oogonia mostly with a paragynous antheridium. It has semi-papillate, caducous sporangia with variable pedicels, but it does not have hyphal swelling. Analyses of ITS, CO1, beta-tubulin and NADH1 sequences revealed that P. aquimorbida is closely related to P. hydropathica, P. irrigata and P. parsiana, and P. taxon 'aquatilis' is related to P. multivesiculata. The optimum temperature for culture growth is 30 and 20 C for E aquimorbida and P. taxon 'aquatilis' respectively. Both taxa were pathogenic to rhododendron plants and caused root discoloration, pale leaves, wilting, tip necrosis and dieback. Their plant biosecurity risk also is discussed.

Irrigation water recycling is an increasingly important practice in agriculture in the context of diminishing water supply and the regulatory requirements in some parts of the world. This practice potentially accumulates and disseminates plant pathogens including Phytophthora species that pose a great threat to agriculture and forest ecosystems. Despite a high economic importance of Phytophthora species, the current understanding of their aquatic ecology is very limited. Therefore, a study was conducted to investigate the distribution and diversity of Phytophthora species in an irrigation reservoir of a commercial nursery in eastern Virginia over two consecutive winters. Multiple baits were deployed in surface water at a run-off entrance, 20, 40, 60 and 80 m from the entrance and near the pump inlet and at various depths at the 20-m station. Ten different Phytophthora species were detected in this study that included P. citrophthora, P. gonapodyides, P. hydropathica, P. inundata, P. irrigata, P. megasperma, P. pini, P. polonica, P. syringae and P. tropicalis. Phytophthora recovery declined through the winters from November to March. It also declined with distance from the run-off entrance. These results suggest that water decontamination during winter irrigation events is required at this nursery and possibly in the nurseries from the southern part of the United States. The placement of the pump inlet away from run-off entrance may be a viable strategy to reduce the crop health risk.

Phytophthora pini was named by Leonian in 1925, but this species was largely ignored until 1956 and then merged with P. citricola by Waterhouse in 1963. This study compared the ex-type and ex-authentic cultures of these two species with isolates of P. plurivora and the P. citricola subgroups Cil I and III reported previously. Examination of these isolates revealed that the ex-type culture of P. pini is identical to P. citricola I. Phytophthora pini Leonian therefore is resurrected to distinct species status and redescribed here with a Latin description, replacing P. citricola I. Molecular, physiological and morphological descriptions of this species are presented. The molecular description includes DNA sequences of five nuclear and mitochondrial regions as well as PCR-SSCP fingerprints. The relationship among the above species and other species recently segregated from the P. citricola complex also is discussed.

Oomycetes attack a huge variety of economically and ecologically important plants. These pathogens release, detect and respond to signal molecules to coordinate their communal behaviors including the infection process. When signal molecules are present at or above threshold level, single zoospores can infect plants. However, at the beginning of a growing season population densities of individual species are likely below those required to reach a quorum and produce threshold levels of signal molecules to trigger infection. It is unclear whether these molecules are shared among related species and what their chemistries are.